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ACS Synthetic biology v.6 no.1, 2017년, pp.94 - 107   SCIE
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CRISPR Perturbation of Gene Expression Alters Bacterial Fitness under Stress and Reveals Underlying Epistatic Constraints

Otoupal, Peter B. (Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado 80309, ); Erickson, Keesha E. ( Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado 80309, ); Escalas-Bordoy, Antoni ( Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado 80309, ); Chatterjee, Anushree ( Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado 80309, );
  • 초록  

    The evolution of antibiotic resistance has engendered an impending global health crisis that necessitates a greater understanding of how resistance emerges. The impact of nongenetic factors and how they influence the evolution of resistance is a largely unexplored area of research. Here we present a novel application of CRISPR-Cas9 technology for investigating how gene expression governs the adaptive pathways available to bacteria during the evolution of resistance. We examine the impact of gene expression changes on bacterial adaptation by constructing a library of deactivated CRISPR-Cas9 synthetic devices to tune the expression of a set of stress-response genes in Escherichia coli . We show that artificially inducing perturbations in gene expression imparts significant synthetic control over fitness and growth during stress exposure. We present evidence that these impacts are reversible; strains with synthetically perturbed gene expression regained wild-type growth phenotypes upon stress removal, while maintaining divergent growth characteristics under stress. Furthermore, we demonstrate a prevailing trend toward negative epistatic interactions when multiple gene perturbations are combined simultaneously, thereby posing an intrinsic constraint on gene expression underlying adaptive trajectories. Together, these results emphasize how CRISPR-Cas9 can be employed to engineer gene expression changes that shape bacterial adaptation, and present a novel approach to synthetically control the evolution of antimicrobial resistance. Graphic Abstract ACS Electronic Supporting Info


  • 주제어

    CRISPR .   transcriptome .   adaptive evolution .   epistasis .   antibiotic resistance.  

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